Chemical grafting of molecularly imprinted homopolymers to the surface of microplates. Application of artificial adrenergic receptor in enzyme-linkedassay for beta-agonists determination

A technique for coating of microplate weds with a molecularly imprinted pol ymer (MTP), specific for epinephrine, is presented. 3-Aminophenylboronic ac id was polymerized in the presence of epinephrine using oxidation of the mo nomer by ammonium persulfate. This process resulted in the grafting of a th in polymer layer onto the polystyrene surface of the microplates. The polym er affinity was determined by an enzyme-linked assay using a conjugate of h orseradish peroxidase and norepinephrine (HRP-N). It was found that imprint ing resulted in increased affinity of the polymer toward HRP-N and epinephr ine. Influence of the buffer pH and concentration on the polymer affinity w as analyzed. It was shown that the MIP-coated microplates could be used for assay development and drug screening. The high stability of the polymers a nd good reproducibility of the measurements make MIP coating an attractive alternative to traditional antibodies or receptors, used in ELISA.